Reproductive Medicine Center, Department of Obstetrics and Gynecology, The Second Xiangya Hospital, Central South University, Changsha, Hunan, China.
Shandong Binzhou Animal Science & Veterinary Medicine Academy, Binzhou, Shandong, China.
Microb Pathog. 2021 Jun;155:104882. doi: 10.1016/j.micpath.2021.104882. Epub 2021 Apr 10.
Psittacosis is a zoonotic disease caused by Chlamydia psittaci (C. psittaci), leading to high risk for animal industry and human health. Lack of reliable commercial kits and effective vaccines is hampering control of C. psittaci infection. Polymorphic outer membrane protein Gs (PmpGs) are enriched in diverse C. psittaci, and its role are unclear during C. psittaci infection. In the present study, pmp20G gene was cloned into pET-28a vector and then the constructed plasmid was transferred into Escherichia coli Rossetta (DE3). After denaturation and renaturation, the recombinant Pmp20G-N was identified by SDS-PAGE and Western blot. Afterwards Pmp20G-N was used as the coating antigen to develop an indirect ELISA (I-ELISA) assay. Both the specificity and sensitivity of Pmp20G-N ELISA were 100%, while the MOMP-ELISA had 93.65% sensitivity and 98.94% specificity, respectively. The concordance between MOMP-ELISA and Pmp20G-N ELISA assay was 98.1%. Hence, Pmp20G-N ELISA has the potential to be a diagnostic antigen for detection C. psittaci antibody. However, further studies are needed to be done for differentiating C. psittaci from Chlamydia spp. and other C.psittaci-specific serovars using Pmp20G-N ELISA.
鹦鹉热是由鹦鹉热衣原体（C. psittaci）引起的一种人畜共患病，对动物产业和人类健康构成高风险。缺乏可靠的商业试剂盒和有效的疫苗阻碍了对 C. psittaci 感染的控制。多态性外膜蛋白 Gs（PmpGs）在各种 C. psittaci 中丰富存在，但其在 C. psittaci 感染期间的作用尚不清楚。在本研究中，pmp20G 基因被克隆到 pET-28a 载体中，然后将构建的质粒转入大肠杆菌 Rosetta（DE3）。变性和复性后，通过 SDS-PAGE 和 Western blot 鉴定重组 Pmp20G-N。然后将 Pmp20G-N 用作包被抗原来开发间接 ELISA（I-ELISA）检测。Pmp20G-N ELISA 的特异性和敏感性均为 100%，而 MOMP-ELISA 的敏感性和特异性分别为 93.65%和 98.94%。MOMP-ELISA 和 Pmp20G-N ELISA 检测之间的一致性为 98.1%。因此，Pmp20G-N ELISA 有可能成为检测 C. psittaci 抗体的诊断抗原。然而，需要进一步研究使用 Pmp20G-N ELISA 来区分 C. psittaci 与衣原体属和其他 C.psittaci 特异性血清型。
